RUNX3 regulates FAP to influence the proliferation of mouse lung primary fibroblasts
10.19405/j.cnki.issn1000–1492.2026.04.003
- VernacularTitle:RUNX3通过调控FAP对小鼠肺原代成纤维细胞增殖的影响
- Author:
Junbo YOU
1
;
Xianchen WANG
1
;
Hui LING
1
;
Jiahao FAN
1
;
Qi CHEN
2
;
Hui TAO
2
;
Jiming SHA
1
Author Information
1. Dept of Thoracic Surgery, The Second Affiliated Hospital of Anhui Medical University， Hefei 230601
2. Dept of Anesthesiology and Perioperative Medicine, The Second Affiliated Hospital of Anhui Medical University， Hefei 230601
- Publication Type:Journal Article
- Keywords:
RUNX3;
primary pulmonary fibroblasts;
fibroblast activation protein;
proliferation;
pulmonary fibrosis
- From:
Acta Universitatis Medicinalis Anhui
2026;61(4):606-611
- CountryChina
- Language:Chinese
-
Abstract:
ObjectiveTo investigate the role of runt-related transcription factor 3 （RUNX3） in transforming growth factor-β1 （TGF-β1）-induced activation of mouse primary pulmonary fibroblasts （PFs）， and its effects on fibroblast activation protein （FAP） expression， cell proliferation， and collagen synthesis. MethodsPFs were isolated from C57BL/6 mice and cultured. A RUNX3 knockdown model was established using small interfering RNA （siRNA）. Cells were assigned to the control group （Control）， TGF-β1-treated group （TGF-β1）， negative control group （TGF-β1+siRNA-NC）， and RUNX3-silenced group （TGF-β1+si-RUNX3）. In addition， a RUNX3 overexpression rescue experiment was performed based on TGF-β1 stimulation. Protein and mRNA levels of RUNX3， FAP， and typeⅠcollagen （COL1A1） were measured by Western blot and reverse transcription quantitative real-time PCR （RT-qPCR）. Cell proliferation was assessed using CCK-8 and EdU assays. Co-expression of COL1A1 and FAP was examined by double immunofluorescence staining. ResultsCompared with the Control group， RUNX3， FAP， and COL1A1 expression levels were upregulated in PFs in the TGF-β1 group （P<0.01）. The CCK-8 assay showed that the absorbance value was reduced in the RUNX3 knockdown group compared with the negative control group （P<0.01）. Consistently， the EdU assay demonstrated a lower proportion of EdU-positive cells in the RUNX3 knockdown group than in the negative control group （P<0.01）. Immunofluorescence double staining revealed decreased fluorescence intensities of COL1A1 and FAP in the RUNX3 knockdown group relative to the negative control. Under RUNX3 overexpression conditions， these fluorescence signals exhibited a partial rebound （P<0.01）. ConclusionRUNX3 in TGF-β1-induced PFs may promote cell proliferation and collagen synthesis by positively regulating FAP expression. Targeting the RUNX3/FAP axis may represent a potential therapeutic strategy for pulmonary fibrosis.
